CN102040531A - Method for extracting L-isoleucine - Google Patents
Method for extracting L-isoleucine Download PDFInfo
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- CN102040531A CN102040531A CN201010557488XA CN201010557488A CN102040531A CN 102040531 A CN102040531 A CN 102040531A CN 201010557488X A CN201010557488X A CN 201010557488XA CN 201010557488 A CN201010557488 A CN 201010557488A CN 102040531 A CN102040531 A CN 102040531A
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Abstract
The invention discloses a method for extracting L-isoleucine, belonging to the field of biochemical engineering. The method disclosed by the invention comprises the following steps of: flocculating an L-isoleucine fermentation liquid, and micro-filtering the flocculated clear solution by using a ceramic membrane with pore diameter of 50nm and molecular weight of 30KD; adsorbing the filtration liquid obtained by micro-filtration by using strong acid cation exchange resin, eluting the strong acid cation exchange resin by using 0.2-0.6mol/L ammonia water or sodium hydroxide, collecting an elution liquid A from the starting of elution to the moment when the pH value of the elution liquid is 7-8, and collecting an elution liquid B from the moment when the pH value of the elution liquid is 7-8 to the moment when the pH value of the elution liquid is 9.5-10.5; concentrating, filtering and decolorizing the elution liquid A, crystallizing to obtain L-isoleucine crystals, and repeating ion exchange adsorption on the elution liquid B. The method can be used for effectively removing impurities in the filtration liquid and the elution liquids, increases the extraction rate and the quality of L-isoleucine and can be widely applied to the production of the L-isoleucine.
Description
Technical field
The present invention relates to biological chemical field, a kind of method of the L-of extraction Isoleucine specifically is provided.
Background technology
Isoleucine, formal name used at school 2-amino-3 methylvaleric acid, the another name L-iLeu is the wherein a kind of of 20 kinds of primary amino acids, almost is present in the structure of all proteins.Isoleucine has 2 unsymmetrical carbons, so there is the diastereomer of 4 kinds of steric isomers and 2 L-Isoleucines in it, but only there is one type of L-Isoleucine in occurring in nature.Because the L-Isoleucine can not synthesize in human body, so it is an essential amino acid, is referred to as branched-chain amino acid with L-leucine, L-Xie Ansuan, because of its special 26S Proteasome Structure and Function, has vital role in the human life metabolism.
In the world, L-Isoleucine manufacturer mainly contains 4 companies such as Japanese aginomoto, consonance fermentation, the pharmacy of limit, field and German Degussa, all adopts fermentation method explained hereafter L-Isoleucine.Wherein, Japan occupies first place in the world on L-Isoleucine output, quality and science and technology, and its L-Isoleucine acid production rate is 35-50g/L, and extraction yield is 70%.At home, manufacturer mainly contains the suitable medicine group company in Hubei, Three Gorges, Yichang pharmaceutcal corporation, Ltd, the Wuxi extra large amino acid of crystalline substance company, Nanning An Litai medicine company limited liability company etc.Simultaneously domestic also have many reports and patent to ion exchange adsorption extraction L-Isoleucine, for example, coming rosy clouds and Qinghua LIU to report on Shenyang Pharmaceutical University's journal, Wuxi Light Industry Univ.'s journal respectively utilizes ion exchange method to extract the technology of L-Isoleucine; Chinese patent CN200510123082.X discloses ion exchange method is extracted the L-Isoleucine from fermented liquid process for cleanly preparing.But the extraction yield of the L-Isoleucine in these reports has only 50～60%, and quality is also lower.
Cause the extraction yield and the lower major cause of quality of L-Isoleucine to be, present technology is generally taked the method for disposable collecting elutriant in the ion-exchange absorption stage, but prolongation along with elution time, assorted aminoacids content in the elutriant can increase gradually, and this extraction and quality to the L-Isoleucine has caused influence.In addition, present filtration process can't be removed tropina to greatest extent, and the crystallization processes of present stage also can't gather in the crops L-Isoleucine crystal efficiently, and these also are to cause one of lower factor of L-Isoleucine extraction yield and quality.
Summary of the invention
In view of this, the invention provides a kind of method of the L-of extraction Isoleucine, this method has solved effectively in the ion exchange method because the disposable collecting elutriant, causes that assorted amino acid increases the problem that influence extraction of L-Isoleucine in the elutriant, has improved the extraction yield and the quality of L-Isoleucine.
The invention provides a kind of method of the L-of extraction Isoleucine, comprising:
Step 1, with L-isoleucine fermentation liquid flocculation, the settled solution after the flocculation is that 50nm, molecular weight are the ceramic membrane micro-filtration of 30KD with the aperture;
Gained filtrate is adsorbed with strong acidic ion resin behind step 2, the micro-filtration, use the ammoniacal liquor of 0.2～0.6mol/L or sodium hydroxide to the strong acidic ion resin wash-out then, collecting that wash-out begins to elutriant pH value is 7～8 o'clock A elutriant, and collection elutriant pH value is that 7～8 to elutriant pH values are 9.5～10.5 o'clock B elutriant;
Step 3, described A elutriant concentrate, filter, after the decolouring, crystallization, obtain L-Isoleucine crystal;
Step 4, B elutriant adsorb with strong acidic ion resin, use the ammoniacal liquor of 0.2～0.6mol/L or sodium hydroxide to the strong acidic ion resin wash-out then, collect wash-out begin to elutriant pH value be that 7～8 o'clock elutriant concentrates, filters, acquisition L-Isoleucine crystal after the decolouring, crystallization;
Step 5, collection elutriant pH value are that 7～8 to elutriant pH values are 9.5～10.5 o'clock B elutriant repeating step 4.
Particular flow sheet is referring to Fig. 1.
Wherein, L-isoleucine fermentation liquid is to obtain after producing bacterial strain and fermenting raw materials by industrial L-Isoleucine commonly used.
At filtration stage, existing technology is directly generally to filter after the flocculation, so just makes the repeatedly circulation of L-isoleucine fermentation liquid through the film system, and the thalline of flocculation is smashed, and has influenced the effect of flocculation, makes to have tropina in the filtrate.And the present invention adopts flocculation and ceramic membrane micro-filtration process combined, and the settled solution micro-filtration the while is got flocculation when micro-filtration after has reduced the tropina content of filtrate greatly.
Wherein, the concentration ratio of the filtrate of described micro-filtration and L-isoleucine fermentation liquid is 1: 6～10, described micro-filtration is for being 0.2～0.8Mpa at working pressure, crossing film temperature is 40～60 ℃, add the dialysis water yield and be micro-filtration under 30～50% the operational requirement of L-isoleucine fermentation liquid, described flocculation is flocculated for poly aluminium chloride, chitosan or the positive poly-propionic acid amide with 0.1～4% addition.In the present invention, all per-cents all are meant mass percent.
In the ion-exchange absorption stage, the present invention adopts the method for Fractional Collections elutriant, and the elutriant of different steps is done different treatment.The present invention detects with the elutriant of high performance liquid chromatography to different steps, detected result show from wash-out begin to elutriant pH value be 7～8 o'clock, elutriant L-isoleucine content greater than 0.2% and assorted aminoacids content less than 0.15%, its assorted aminoacids content extracted the later stage of L-Isoleucine does not have influence, so this stepwise elution liquid of name is the A elutriant and collects, directly sends into subsequent processing.Owing to adopt ammoniacal liquor or sodium hydroxide to come wash-out, along with constantly carrying out of wash-out, the pH value of elutriant can be increasing, it is that 7～8 to elutriant pH values are 9.5～10.5 o'clock that detected result shows from elutriant pH value, elutriant L-isoleucine content greater than 0.2% and assorted aminoacids content be 0.15～0.5%, naming this stepwise elution liquid is B elutriant and collection, its contained assorted amino acid exceeds standard slightly, need carry out ion-exchange absorption once more, the quality that so not only guarantees the A elutriant that collect next time is higher, can avoid simultaneously loss, guarantee the extraction yield and the quality of L-Isoleucine elutriant.
Wherein, described assorted amino acid is meant all amino acid of other except that the L-Isoleucine in the elutriant, and described elution speed is 0.2～0.5m/s, and described strong acidic ion resin is 732 types, JK008 type or JK006 type resin.
In other operation stages, the present invention changes original decolouring earlier back enrichment process, adopts the method that concentrates rear decoloring earlier, effectively reduces the consumption of gac.Described A elutriant separates after concentrating, obtain comprising the solid and the mother liquor of L-Isoleucine, most of impurity enters into mother liquor, and the pigment in the solid and other impurity will reduce, not only can reduce the consumption of gac when solid dissolves decolouring more like this, also improve the quality of product.
The method that the present invention adopts gradient cooling, hangs down the incubation crystalline substance at crystallisation stage to-10～8 ℃, is educated brilliant 2～8h with the 12h gradient cooling then.Thick crystal is beneficial to separating, washing and quality is better, and tiny crystal runs off in separation easily.If the use fast cooling, crystal does not have process of growth, can produce a large amount of fine crystals, and thisly separates out too fast crystal and carry impurity easily secretly, can reduce the extraction yield and the quality of L-Isoleucine.Therefore, the present invention uses gradient cooling control crystalline growth velocity, makes crystal slowly be grown to thick crystal, and educates crystalline substance at low temperatures and can further reduce the solubleness of L-Isoleucine in water, is beneficial to and separates out more crystal.
In addition, the method for the invention also comprises the processing of classifying of the raffinate after the described crystallization, is specially:
When the assorted aminoacids content of raffinate less than 4% and sulphate content less than 1% the time, the 3 described decolourings of raffinate repeating step, crystallization; When the assorted aminoacids content of raffinate greater than 4% or sulphate content greater than 1% the time, raffinate repeating step 3 is described to be concentrated, filtration, decolouring, crystallization.
The present invention can effectively remove the impurity in filtrate and the elutriant, improves the extraction yield and the quality of L-Isoleucine, saves cost, helps the conservation of nature environment, can be widely used in the production of L-Isoleucine.
Description of drawings
Figure 1 shows that the method flow diagram of extraction L-Isoleucine of the present invention.
Embodiment
The invention discloses a kind of method of the L-of extraction Isoleucine, those skilled in the art can use for reference this paper content, suitably improve processing parameter and realize.Special needs to be pointed out is that all similarly replace and change apparent to those skilled in the art, they all are regarded as being included in the present invention.Method of the present invention is described by preferred embodiment, and the related personnel obviously can change or suitably change and combination methods and applications as herein described in not breaking away from content of the present invention, spirit and scope, realizes and use the technology of the present invention.
According to the present invention, described a kind of method of extracting the L-Isoleucine comprises:
Step 1, with L-isoleucine fermentation liquid flocculation, the settled solution after the flocculation is that 50nm, molecular weight are the ceramic membrane micro-filtration of 30KD with the aperture;
Gained filtrate is adsorbed with strong acidic ion resin behind step 2, the micro-filtration, use the ammoniacal liquor of 0.2～0.6mol/L or sodium hydroxide to the strong acidic ion resin wash-out then, collecting that wash-out begins to elutriant pH value is 7～8 o'clock A elutriant, and collection elutriant pH value is that 7～8 to elutriant pH values are 9.5～10.5 o'clock B elutriant;
Step 3, described A elutriant concentrate, filter, after the decolouring, crystallization, obtain L-Isoleucine crystal, described B elutriant repeating step 2.
Wherein, the concentration ratio of the filtrate of described micro-filtration and L-isoleucine fermentation liquid is 1: 6～10, described micro-filtration is for being 0.2～0.8Mpa at working pressure, crossing film temperature is 40～60 ℃, add the dialysis water yield and be micro-filtration under 30～50% the operational requirement of L-isoleucine fermentation liquid, described flocculation is flocculated for poly aluminium chloride, chitosan or the positive poly-propionic acid amide with 0.1～4% addition.
Wherein, described elution speed is 0.2～0.5m/s, and described strong acidic ion resin is 732 types, JK008 type or JK006 type resin; Described crystallization is extremely-10～8 ℃ of gradient coolings, educates brilliant 2～8h then.
In addition, the method for the invention also comprises the processing of classifying of the raffinate after the described crystallization, is specially:
When the assorted aminoacids content of raffinate less than 4% and sulphate content less than 1% the time, the 3 described decolourings of raffinate repeating step, crystallization; When the assorted aminoacids content of raffinate greater than 4% or sulphate content greater than 1% the time, raffinate repeating step 3 is described to be concentrated, filtration, decolouring, crystallization.
Through about the L-Isoleucine extraction rate reached to 70% that the method for the invention is extracted, its percentage composition detects through high performance liquid chromatography and reaches more than 98.5%, shows that the method for the invention can improve the extraction yield and the quality of L-Isoleucine.
In addition, the present invention also detects the wet solid content of tropina in the filtrate, and the result is 0, and adopts the general method filtrate filtered, the wet solid content of its tropina is 0.3%, has proved absolutely that present method more can effectively remove tropina than general method.
Below in conjunction with embodiment, further set forth the present invention.
Embodiment 1: the method for the invention is extracted the L-Isoleucine
1, extracting method
Through the L-isoleucine fermentation liquid that the enzyme that goes out is handled, regulate fermented liquid pH to 3 with hydrochloric acid, the addition by 0.1% adds chitosan, keeps 20 ℃ of flocculation temperature, and stirs 30min.The employing aperture is that 50nm, molecular weight are that 30KD, area are 117m
2The settled solution micro-filtration of ceramic membrane after to flocculation, controlling the ceramic membrane working pressure is 0.2Mpa, crosses 40 ℃ of film temperatures, adding the dialysis water yield is 30% (volume ratio) of fermented liquid, the concentration ratio of filtrate and L-isoleucine fermentation liquid is 1: 6.Filtrate is directly adsorbed on 732 type strong acid type cationic resins, and upper prop speed is 0.2m/s.Adsorb saturated after, adopt washed with de-ionized water resin impurity.Adopt 0.2mol/L ammoniacal liquor to carry out wash-out then, elution speed adopts 0.2m/s, collects by wash-out to begin A elutriant to elutriant pH 7; Collection begins B elutriant to elutriant pH 9.5 by elutriant pH 7.
Described B elutriant is sent into the ion-exchange absorption stage once more carry out next one circulation.Described A elutriant enters the single-action crystallizer and concentrates, vacuum tightness≤0.92Mpa, and temperature is controlled at 60 ℃, when the A elutriant after concentrating is 1: 6 with the volume ratio that concentrates preceding A elutriant, is cooled to 0 ℃ and puts jar.With the A elutriant Plate Filtration after concentrating, when feed pressure＞0.3Mpa, stop charging, adopt the 0.3Mpa air to carry out liftout till anhydrous oozing, obtain filter cake.According to filter cake content, adopt deionized water to dissolve then by L-Isoleucine pure 4%.Be warming up to 60 ℃, throw charcoal by 0.5～2%.Bleaching time 1 hour, middle graded is thrown charcoal, until the crude product transmittance greater than 99%.
With the solution evaporation after the decolouring, 60 ℃ of control vaporization temperatures, vacuum tightness≤0.92Mpa stops blowing when the liquor capacity ratio reaches 1: 5 before evaporating back solution and evaporating.Adopt gradient cooling, reduced to-10 ℃ in 12 hours, per hour cooling extent is no more than 6 ℃, and educates under this temperature brilliant 2 hours.After educating brilliant the end, centrifugal to whizzer from educating brilliant jar blowing, till not having raffinate and flowing out, the centre can be according to the suitable wash water of feed liquid situation, and the gained crystal is the L-Isoleucine, and the gained raffinate is done following processing according to feed liquid quality situation:
When the assorted aminoacids content of raffinate less than 4% and sulphate content less than 1% the time, raffinate is sent into bleaching process, enter next circulation; When the assorted aminoacids content of raffinate greater than 4% or sulphate content greater than 1% the time, raffinate is sent into enrichment process, enter next circulation.
2, product detects
Statistics L-Isoleucine product extraction rate is 71.2%; With high performance liquid chromatography product is carried out content detection, the L-isoleucine content is 98.7%.Detected result shows that the method for the invention has improved L-Isoleucine extraction yield, and quality product reaches higher level simultaneously.
Embodiment 2: the method for the invention is extracted the L-Isoleucine
1, extracting method
Through the L-isoleucine fermentation liquid that the enzyme that goes out is handled, regulate fermented liquid pH to 3 with hydrochloric acid, the addition by 4% adds aluminum chloride, keeps 70 ℃ of flocculation temperature, and stirs 120min.The employing aperture is that 50nm, molecular weight are that 30KD, area are 117m
2The settled solution micro-filtration of ceramic membrane after to flocculation, controlling the ceramic membrane working pressure is 0.8Mpa, crosses 60 ℃ of film temperatures, adding the dialysis water yield is 50% (volume ratio) of fermented liquid, the concentration ratio of filtrate and L-isoleucine fermentation liquid is 1: 10.Filtrate is directly adsorbed on JK008 type strong acid type cationic resin, and upper prop speed is 1m/s.Adsorb saturated after, adopt washed with de-ionized water resin impurity.Adopt 0.6N sodium hydroxide to carry out wash-out then, elution speed adopts 0.5m/s, collects by wash-out to begin A elutriant to elutriant pH 8; Collection begins B elutriant to elutriant pH 10.5 by elutriant pH 8.
Described B elutriant is sent into the ion-exchange absorption stage once more carry out next one circulation.Described A elutriant enters the single-action crystallizer and concentrates, vacuum tightness≤0.92Mpa, and temperature is controlled at 80 ℃, when the volume ratio of A elutriant after concentrating and initial A elutriant is 1: 8, is cooled to 30 ℃ and puts jar.With the A elutriant Plate Filtration after concentrating, when feed pressure＞0.3Mpa, stop charging, adopt the 0.3Mpa air to carry out liftout till anhydrous oozing, obtain filter cake.According to filter cake content, adopt deionized water to dissolve then by L-Isoleucine pure 4%.Be warming up to 60 ℃, throw charcoal by 0.5～2%.Bleaching time 1 hour, middle graded is thrown charcoal, until the crude product transmittance reach＞99%.
With the solution evaporation after the decolouring, 60 ℃ of control vaporization temperatures, vacuum tightness≤0.92Mpa stops blowing when the liquor capacity ratio reaches 1: 5 before evaporating back solution and evaporating.Adopt gradient cooling, reduced to 8 ℃ in 12 hours, per hour cooling extent is no more than 5 ℃, and educates under this temperature brilliant 8 hours.After educating brilliant the end, centrifugal to whizzer from educating brilliant jar blowing, till not having raffinate and flowing out, the centre can be according to the suitable wash water of feed liquid situation, and the gained crystal is the L-Isoleucine, and the gained raffinate is done following processing according to feed liquid quality situation:
When the assorted aminoacids content of raffinate less than 4% and sulphate content less than 1% the time, raffinate is sent into bleaching process, enter next circulation; When the assorted aminoacids content of raffinate greater than 4% or sulphate content greater than 1% the time, raffinate is sent into enrichment process, enter next circulation.
2, product detects
Statistics L-Isoleucine product extraction rate is 72.4%; With high performance liquid chromatography product is carried out content detection, the L-isoleucine content is 98.1%.Detected result shows that the method for the invention has improved L-Isoleucine extraction yield, and quality product reaches higher level simultaneously.
Embodiment 3: the method for the invention is extracted the L-Isoleucine
1, extracting method
Through the L-isoleucine fermentation liquid that the enzyme that goes out is handled, regulate fermented liquid pH to 3 with hydrochloric acid, the addition by 4% adds aluminum chloride, keeps 45 ℃ of flocculation temperature, and stirs 60min.The employing aperture is that 50nm, molecular weight are that 30KD, area are 117m
2The settled solution micro-filtration of ceramic membrane after to flocculation, controlling the ceramic membrane working pressure is 0.5Mpa, crosses 50 ℃ of film temperatures, adding the dialysis water yield is 40% (volume ratio) of fermented liquid, the concentration ratio of filtrate and L-isoleucine fermentation liquid is 1: 8.Filtrate is directly adsorbed on JK006 type strong acid type cationic resin, and upper prop speed is 0.6m/s.Adsorb saturated after, adopt washed with de-ionized water resin impurity.Adopt 0.4N ammoniacal liquor to carry out wash-out then, elution speed adopts 0.3m/s, collects by wash-out to begin A elutriant to elutriant pH 8; Collection begins B elutriant to elutriant pH10.5 by elutriant pH 8.
Described B elutriant is sent into the ion-exchange absorption stage once more carry out next one circulation.Described A elutriant enters the single-action crystallizer and concentrates, vacuum tightness≤0.92Mpa, and temperature is controlled at 70 ℃, when the volume ratio of A elutriant after concentrating and initial A elutriant is 1: 7, is cooled to 15 ℃ and puts jar.With the A elutriant Plate Filtration after concentrating, when feed pressure＞0.3Mpa, stop charging, adopt the 0.3Mpa air to carry out liftout till anhydrous oozing, obtain filter cake.According to filter cake content, adopt deionized water to dissolve then by L-Isoleucine pure 4%.Be warming up to 60 ℃, throw charcoal by 0.5～2%.Bleaching time 1 hour, middle graded is thrown charcoal, until the crude product transmittance reach＞99%.
With the solution evaporation after the decolouring, 60 ℃ of control vaporization temperatures, vacuum tightness≤0.92Mpa stops blowing when the liquor capacity ratio reaches 1: 5 before evaporating back solution and evaporating.Adopt gradient cooling, reduced to 0 ℃ in 12 hours, per hour cooling extent is no more than 5 ℃, and educates under this temperature brilliant 5 hours.After educating brilliant the end, centrifugal to whizzer from educating brilliant jar blowing, till not having raffinate and flowing out, the centre can be according to the suitable wash water of feed liquid situation, and the gained crystal is the L-Isoleucine, and the gained raffinate is done following processing according to feed liquid quality situation:
When the assorted aminoacids content of raffinate less than 4% and sulphate content less than 1% the time, raffinate is sent into bleaching process, enter next circulation; When the assorted aminoacids content of raffinate greater than 4% or sulphate content greater than 1% the time, raffinate is sent into enrichment process, enter next circulation.
2, product detects
Statistics L-Isoleucine product extraction rate is 73.3%; With high performance liquid chromatography product is carried out content detection, the L-isoleucine content is 97.7%.Detected result shows that the method for the invention has improved L-Isoleucine extraction yield, and quality product reaches higher level simultaneously.
Embodiment 4: the method for the invention is gone the tropina effect detection
Tropina content in the L-isoleucine fermentation liquid is represented with wet solid content usually.The detection method of wet solid content draws its content in fermented liquid for centrifugal mode separating thallus albumen thereby measure wet proteic quality then.
The filtrate of the present invention after to micro-filtration among the embodiment 1 to 3 is carried out the wet solid content of tropina and is detected, and as a comparison, the filtrate of adopting the general method direct filtration and unfiltered fermented liquid is also carried out the wet solid content of tropina detect, and concrete outcome is referring to table 1.
Table 1 tropina content
| Do not filter | General method | Embodiment 1 | Embodiment 2 | Embodiment 3 | |
| The tropina solid content that wets | 15％ | 0.3％ | ?0 | ?0 | ?0 |
The result shows, L-isoleucine fermentation liquid is behind process the method for the invention micro-filtration, and the wet solid content of tropina is 0, and adopts the filtrate after the general method direct filtration, the wet solid content of its tropina is 0.3%, and the wet solid content of unfiltered fermented liquid tropina is 15%.Experimental data shows that the method for the invention can remove tropina in the fermented liquid effectively, eliminates its influence to L-Isoleucine extraction yield and quality.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (8)
1. a method of extracting the L-Isoleucine is characterized in that, comprising:
Step 1, with L-isoleucine fermentation liquid flocculation, the settled solution after the flocculation is that 50nm, molecular weight are the ceramic membrane micro-filtration of 30KD with the aperture;
Gained filtrate is adsorbed with strong acidic ion resin behind step 2, the micro-filtration, use the ammoniacal liquor of 0.2～0.6mol/L or sodium hydroxide to the strong acidic ion resin wash-out then, collecting that wash-out begins to elutriant pH value is 7～8 o'clock A elutriant, and collection elutriant pH value is that 7～8 to elutriant pH values are 9.5～10.5 o'clock B elutriant;
Step 3, A elutriant concentrate, filter, after the decolouring, crystallization, obtain L-Isoleucine crystal;
Step 4, B elutriant adsorb with strong acidic ion resin, use the ammoniacal liquor of 0.2～0.6mol/L or sodium hydroxide to the strong acidic ion resin wash-out then, collect wash-out begin to elutriant pH value be that 7～8 o'clock elutriant concentrates, filters, acquisition L-Isoleucine crystal after the decolouring, crystallization;
Step 5, collection elutriant pH value are that 7～8 to elutriant pH values are 9.5～10.5 o'clock B elutriant repeating step 4.
2. according to the described method of claim 1, it is characterized in that described flocculation is poly aluminium chloride, chitosan or positive poly-propionic acid amide flocculation with 0.1～4% addition.
3. according to the described method of claim 1, it is characterized in that the concentration ratio of the filtrate of described micro-filtration and L-isoleucine fermentation liquid is 1: 6～10.
4. according to the described method of claim 3, it is characterized in that described micro-filtration is for being 0.2～0.8Mpa at working pressure, crossing film temperature is 40～60 ℃, adds the dialysis water yield and be micro-filtration under 30～50% the operational requirement of L-isoleucine fermentation liquid.
5. according to the described method of claim 1, it is characterized in that described wash-out is the speed wash-out with 0.2～0.5m/s.
6. according to the described method of claim 1, it is characterized in that described strong acidic ion resin is 732 types, JK008 type or JK006 type resin.
7. according to the described method of claim 1, it is characterized in that described crystallization is extremely-10～8 ℃ of gradient coolings, educates brilliant 2～8h then.
8. according to the described method of claim 1, it is characterized in that, also comprise, be specially the processing of classifying of the raffinate after the described crystallization:
When the assorted aminoacids content of raffinate less than 4% and sulphate content less than 1% the time, the 3 described decolourings of raffinate repeating step, crystallization; When the assorted aminoacids content of raffinate greater than 4% or sulphate content greater than 1% the time, raffinate repeating step 3 is described to be concentrated, filtration, decolouring, crystallization.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102505027A (en) * | 2011-12-27 | 2012-06-20 | 开原市天慕生物科技有限公司 | Isoleucine fermenting process |
| CN102643277A (en) * | 2012-04-10 | 2012-08-22 | 常州康丽制药有限公司 | Refining method of ganciclovir |
| CN104450815A (en) * | 2014-11-20 | 2015-03-25 | 河南巨龙生物工程股份有限公司 | Fermentation method for improving yield of isoleucine |
| CN104744277A (en) * | 2015-02-12 | 2015-07-01 | 新疆阜丰生物科技有限公司 | Simulated moving bed and method for extracting three-branch chain amino acid based on simulated moving bed |
| CN106220521A (en) * | 2016-08-11 | 2016-12-14 | 山东阜丰发酵有限公司 | A kind of full film extracts the method for L isoleucine |
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| CN104744277A (en) * | 2015-02-12 | 2015-07-01 | 新疆阜丰生物科技有限公司 | Simulated moving bed and method for extracting three-branch chain amino acid based on simulated moving bed |
| CN107129436A (en) * | 2016-06-27 | 2017-09-05 | 通辽梅花生物科技有限公司 | A kind of isoleucine extraction process |
| CN106278919B (en) * | 2016-08-11 | 2018-01-16 | 山东阜丰发酵有限公司 | A kind of method for preparing L isoleucines |
| CN106278919A (en) * | 2016-08-11 | 2017-01-04 | 山东阜丰发酵有限公司 | A kind of method preparing L isoleucine propylhomoserin |
| CN106220521B (en) * | 2016-08-11 | 2018-01-16 | 山东阜丰发酵有限公司 | A kind of method of full film extraction L isoleucines |
| CN106220521A (en) * | 2016-08-11 | 2016-12-14 | 山东阜丰发酵有限公司 | A kind of full film extracts the method for L isoleucine |
| CN106699587A (en) * | 2016-12-19 | 2017-05-24 | 宜昌三峡制药有限公司 | Isoleucine crystallizing and washing method and device |
| CN109851514A (en) * | 2019-02-24 | 2019-06-07 | 内蒙古拜克生物有限公司 | A kind of isolation and purification method of l-Isoleucine |
| CN109851514B (en) * | 2019-02-24 | 2019-12-13 | 内蒙古拜克生物有限公司 | Separation and purification method of L-isoleucine |
| CN113277955A (en) * | 2021-06-21 | 2021-08-20 | 通辽梅花生物科技有限公司 | Method for extracting L-isoleucine |
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